Therapeutic Antibodies for Treatment and Prophylaxis of Transmittable Viral Diseases

ABSTRACT

The invention provides methods and compositions for the treatment and prevention of a transmittable disease in a subject, such as avians and mammals. The methods and compositions of the invention specifically make use of avian antibodies to the disease to be treated or prevented. Administration of such avian antibodies to a subject has been shown effective for reducing mortality in a population of subjects that are infected, or become infected, with the disease. The invention also provides kits useful for detecting the presence of transmittable diseases in subjects.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation-in part of U.S. patentapplication Ser. No. 12/176,793, filed Jul. 21, 2008, which is acontinuation of U.S. patent application Ser. No. 11/459,832, filed Jul.25, 2006 (now abandoned), which claims priority to Provisional PatentApplication Ser. No. 60/595,652, filed Jul. 25, 2005, all of which areincorporated herein by reference in their entirety.

FIELD OF THE INVENTION

The present invention is directed to compositions and the use thereof inthe treatment and prevention of transmittable diseases, and particularlyviral diseases. The compositions incorporate serum comprising avianantibodies against the transmittable disease, and the compositions canbe used in a variety of subjects, including avians and mammals.

BACKGROUND

Previously known approaches to dealing with epidemiological outbreaks oftransmittable clinical diseases have traditionally focused on threeapproaches: isolation of affected individuals; use of antimicrobialagents, and use of vaccinations. Antimicrobial agents have been usedsuccessfully for treatment once the pathogen has been identified;however, if the microorganism is resistant to the antimicrobial agent,there are limited or no options other than relying on the patient's ownimmune system for recovery or survival (in the case of life-threateninginfections).

Individuals have been routinely protected by vaccinating, or immunizing,against an attenuated bacterial or viral strain where the vaccine hasdemonstrated good efficacy in prior tests. The underlying flaws ofvaccinations are its safety, lack of protection against diverse strainscausing the disease, availability of sufficient supplies of the vaccine,and most importantly, administration of the vaccine in sufficient timeprior to infection to elicit an immune response in the patient againstthe pathogen. Unfortunately, in the event that the population is notvaccinated by the time an outbreak reaches epidemic proportions, avaccination program that requires multiple injections over a significantperiod of time would have very limited effectiveness in protecting thepopulation. In addition, individuals having impaired immunity (i.e., areimmunodeficient) would be unable to generate an effective response.Moreover, given the high cost of a broad vaccination program, thegeneral population has been vaccinated to only a limited number ofpathogens. The rise of numerous emerging infectious diseases and thethreat of bioterrorism acts have significantly elevated thesusceptibility of large populations to a potentially epidemic diseaseoutbreak.

Another approach, which has been referred to as “passive therapeuticimmunity,” to dealing with infection is the use of therapeuticantibodies for the treatment of pathogenic agents that are incurable byantimicrobial agents. Passive therapeutic immunity may also be used forindividuals who have not been previously vaccinated. For example, theuse of therapeutic antibodies has been reported with different degreesof protection against anthrax, biological toxins, brucellosis, Q fever,plague, smallpox, tularemia, viral encephalitides, and viral hemorrhagicfevers. Recent work has focused on the use of monoclonal antibodies,particularly because they can be produced in cell culture in largequantities once the hybridoma cell line is isolated. Alternatively, arecombinant mouse monoclonal antibody can be engineered with humansequences (generally referred to as a “humanized antibody”) and producedin large quantities, albeit at expensive costs that may be prohibitoryfor broad use.

A severe drawback of the use of monoclonal antibodies is that theyrecognize only a single site or epitope on the microorganism, which isnot as effective as polyclonal antibodies that recognize multiple sites.For example, previous testing using anthrax polyclonal sera containingantibodies to several sites demonstrated protective efficacy of thepolyclonal antibodies. However, when the same test was performed usingmonoclonal antibodies, only one of four monoclonal antibodies testedconferred protection. Another limitation of monoclonal antibodytreatment is that monoclonal antibodies offer limited protection topathogens where the epitope is not conservatively maintained, such as apathogen having numerous species or viral pathogens that prone to ahigher mutation frequency.

West Nile Virus is a specific example of a disease where treatment aftercontacting the disease shows little efficacy. Specifically, it isrecognized in the art that there is not yet any experimental evidencethat therapy with immunoglobulin will improve survival or neurologicaloutcome of experimental animals when this therapy is initiated after thedevelopment of the clinical neurological disease. Further, no studies,either prophylactic for protection or post-infection for therapy, havedemonstrated effectiveness of immunoglobulin treatment in animals thatbecome infected by natural transmission of West Nile Virus.

Published U.S. Patent Application 2003/0211110 to Shimoni et al.discloses that hyperimmune sera collected from humans was able tofacilitate the recovery of two immunocompromised patients testedpositive by West Nile Virus upon continuous treatment with antibodydelivered intravenously. In a separate report by Jackson, Can. J.Neurol. Sci., 2004, however, a patient showed no beneficial effect uponsimilar treatment. It is therefore unclear whether the specifiedtreatment alone was responsible for the recovery of the patients, andmore so, if immunosuppression was a key factor required for treatment.

In light of the above, it is clear that further, more effective methodsof treating and preventing infection, particularly by a transmittableviral disease, are needed. The present invention provides pharmaceuticalcompositions and methods of preparation and use thereof that areparticularly beneficial for treating and preventing such infection.

SUMMARY OF THE INVENTION

The present invention relates to the use of avian antibodies,particularly goose antibodies, for treating and preventing infection ina subject by a transmittable viral disease. As such, the invention canprovide specific compositions that are useful in treatment as describedherein, and the invention also can provide specific methods of treatmentas described herein. In particular embodiments, the invention isspecifically useful for reducing mortality in mammals that becomeinfected or are infected with a transmittable viral disease. Theinvention can be practiced in relation to any mammal, particularlyhumans.

In certain embodiments, the invention particularly can provide methodsof treating a mammal infected with a virus in the Flaviviridae family,particularly a virus in the flavivirus genus. Specifically, the methodscan comprise administering to the mammal an amount of a serum comprisingpolyclonal goose antibodies against a virus in the Flaviviridae family.In further embodiments, antibodies from other avians also can be usefulaccording to the invention, such as chicken antibodies.

In specific embodiments, the polyclonal goose antibodies can beantibodies against the very same virus that is infecting the mammalbeing treated according to the invention. The present invention,however, also can provide for treatment wherein the polyclonal gooseantibodies are antibodies against a first virus in the Flaviviridaefamily (or specifically the flavivirus genus) and the virus infectingthe mammal is a second, different virus in the Flaviviridae family (orspecifically the flavivirus genus). Thus, the invention can provide forintra-family or intra-genus treatment (i.e., antibodies against onevirus in a family or genus being effective for treatment of infection bya different virus in the same family or genus). In certain embodiments,the invention particularly can relate to treatment or prophylaxis ofinfection by a flavivirus in the mammalian tick-borne virus group, theseabird tick-borne virus group, the Aroa virus group, the Dengue virusgroup, the Japanese encephalitis virus group, the Kokobera virus group,the Ntaya virus group, the Spondweni virus group, the Yellow fever virusgroup, the Entebbe virus group, the Modoc virus group, or the Rio Bravovirus group.

In other embodiments, the invention particularly can provide methods oftreating a mammal infected with an influenza virus. Specifically, themethods can comprise administering to the mammal an amount of a serumcomprising polyclonal goose antibodies against an influenza virus. Infurther embodiments, antibodies from other avians also can be usefulaccording to the invention.

In specific embodiments, the polyclonal goose antibodies can beantibodies against the very same influenza virus that is infecting themammal being treated according to the invention. The present invention,however, also can provide for treatment wherein the polyclonal gooseantibodies are antibodies against a first influenza virus (orspecifically a virus of the influenzavirus A type) and the virusinfecting the mammal is a second, different influenza virus (orspecifically a virus of the influenzavirus A type). In certainembodiments, the invention particularly can relate to treatment orprophylaxis of infection by a human influenza virus, an avian influenzavirus, a swine influenza virus, an equine influenza virus, a canineinfluenza, or a cat influenza virus.

In further embodiments, the invention can provide methods of treating amammal infected with a virus in a variety of further virus families.Viruses in any of the viral families disclosed herein that affectmammals, particularly humans, can be encompassed by the methods andcompositions of the present invention.

In some embodiments, the serum used in the compositions and methods ofthe invention can be particularly characterized by its surprisingly highneutralization titer as evaluated according to a plaque reduction test.In preferred embodiments, a serum comprising polyclonal goose antibodies(or other avian antibodies) can exhibit a neutralization titer of atleast about 1:500, at least about 1:1000, or even greater, as otherwisedescribed herein.

The invention further can be particularly characterized by the abilityto provide treatment without the requirement of initial actions toprepare a subject for treatment. For example, the invention particularlycan be used in embodiments wherein the mammal being treated with gooseantibodies has not been de-sensitized to goose antibodies prior tocarrying out the invention (i.e., by administering a serum comprisingthe polyclonal goose antibodies against the virus).

In other embodiments, the invention particularly can relate to apharmaceutical composition. For example, such composition can comprise aserum effective for treatment or prophylaxis of a viral infection in amammal arising from a virus in the Flaviviridae family. In suchembodiments, the serum preferably can comprise polyclonal gooseantibodies against a virus in the Flaviviridae family. In particular,the serum can exhibit a neutralization titer for the antibodies of atleast about 1:500 when evaluated according to a plaque reduction test.In another example, a composition according to the invention cancomprise a serum effective for treatment or prophylaxis of a viralinfection in a mammal arising from an influenza virus. In suchembodiments, the serum preferably can comprise polyclonal gooseantibodies against an influenza virus. In particular, the serum canexhibit a neutralization titer for the antibodies of at least about1:500 when evaluated according to a plaque reduction test. Compositionsaccording to the invention can comprise additional components inaddition to the noted serum. For example, the compositions may comprisea pharmaceutically acceptable carrier. Likewise, the compositions may beprovided in kits wherein the composition can be provided in a specificunit volume (e.g., in a vial). Such kits could comprise multiplecontainers (or vials) of the composition, as well as further components,such s instructions for administration of the composition and/ordilutions suitable for providing multiple dosings of the composition.

DETAILED DESCRIPTION OF THE INVENTION

The present inventions now will be described more fully hereinafter withreference to specific embodiments of the invention and particularly tothe various drawing provided herewith. Indeed, the invention may beembodied in many different forms and should not be construed as limitedto the embodiments set forth herein; rather, these embodiments areprovided so that this disclosure will satisfy applicable legalrequirements. As used in the specification, and in the appended claims,the singular forms “a”, “an”, “the”, include plural referents unless thecontext clearly dictates otherwise.

In describing the present invention, various terms and phrases may beused herein, and such terms and phrases will have the same meaningthought the specification.

“Serum” means any fraction of blood serum that contains antibodies andone or more further protein blood components, said protein bloodcomponents comprising at least 30% by weight of the serum.

“Polyclonal antibodies” means a fraction of antibodies isolated from ahost, the fraction comprising at least 80% of the total group ofdifferent antibodies produced by different immune cells in the hosthaving affinity for different antigens on a virus.

“Neutralization titer” means quantification of an antibody that preventsor treats viral infection and subsequent detrimental viral effects,including cell death, in vitro, such as cell cultures including plaqueassays, or in vivo, such as animal testing and clinical treatment.Antibodies that are shown present in a sample by ELISA, Westernanalyses, or like methods may or may not exhibit a neutralization titer.

“Natural transmission” means the transmission of an indigenoustransmittable viral disease that occurs as a result of one or moreanimals being infected in an environment where the animal is freelyexposed to carriers of the transmittable viral disease.

“Naturally occurring strain” or “indigenous strain” means a viral strainthat is present in the natural environment and not having beenreproduced in a laboratory.

“Attenuated” means a viral strain that has been weakened or made lessvirulent.

“Gosling” means any goose that has not reached maturity in terms ofreaching a breeding age.

The treatment of human patients with mammalian antibodies is known tocause strong immunoreactions in non-immunosuppressed human patients. Thetreatment of human patient with non-human, mammalian antibodies likewisecan cause strong immunoreactions. For example, administration of horseantivenin can produce side effects in humans such as severe allergicreactions, and, in extreme cases, death. Therefore, in theever-increasing need for effective treatment and prevention of disease,particularly in relation to viral diseases, an alternative to mammalianantibodies is needed.

Avians are one potential source for antibodies with reduced risk ofimmunoreaction. Even then, the prior art has suggested thatdesensitization is required for cross-species antibody therapies. Thepresent invention has found, however, that avian antibodies fromspecific sources can exhibit fewer or no side effects because they donot activate mammalian complement systems, bind to mammalian rheumatoidfactors, naturally occurring anti-mammalian antibodies, or mammalian Fcreceptors. The present invention thus realizes the ability to useantibodies from specific avian sources to treat or prevent infection bya transmittable viral disease in other avians and mammals.

According to the present invention, it has been found that aviantherapeutic antibodies can be an effective means to protect and treat apopulation of birds in the field by delivery of an effective dose of thetherapeutic antibodies. Moreover, it has been found that aviantherapeutic antibodies can also be an effective means to protect andtreat mammals.

In certain embodiments, the present invention thus provides methods oftreating subjects by administering to the subjects avian antibodiesagainst a transmittable viral disease. Advantageously, in specificembodiments, the present invention allows for treatment wherein thesubject being treated has not been de-sensitized to the avian (e.g.,goose) antibodies prior to treatment via administration of theantibodies. In other words, it can be understood that the subject beingtreated has not been fed or otherwise administered compositionscontaining avian (e.g., goose) antibodies, particularly for the purposeof achieving de-sensitization, prior to treatment by administering theantibodies against the transmittable viral disease. Thus, in methods oftreatment according to the invention, treatment may expressly excludeany step related to de-sensitization of the subject (i.e., mammal) priorto administration of the avian antibodies against the transmittableviral disease.

In specific embodiments, the avian antibodies are goose antibodies. Forsimplicity, the invention may be described herein in relation to geeseand goose antibodies. It is understood, however, that one of skill inthe art armed with the present disclosure may be capable of extendingthe present invention in relation to further avians and avianantibodies, and such are believed to be encompassed by the presentinvention. In particular, the present invention should be recognized asrelating to antibodies obtained, in certain embodiments, from otherwaterfowl, such as ducks, or from turkeys or ostriches. In still furtherembodiments, the invention may be related to antibodies derived fromchickens.

Therefore, according to various embodiments of the present invention,there are provided compositions and methods of use thereof for treatingor preventing infection in a subject by a transmittable viral disease.The invention is particularly characterized by the realization of thetherapeutic ability of high neutralization titer serum to atransmittable viral disease collected from animals that were naturallyinfected with the transmittable viral disease. Alternatively, highneutralization titer sera may be produced by immunization with anattenuated strain of a transmittable viral disease.

Treatment and prevention, according to the invention, are particularlyevidenced by a reduction in the mortality rate in a population ofsubjects to which the serum is administered. Previous attempts to affecttreatment of so-called “previously un-curable diseases” have beenlimited to alleviation or amelioration of symptom associated with thedisease. Moreover, while such previous attempts have alleged prolongingthe survival of infected subjects, such attempts have specificallyfailed to evidence actual reduction in the mortality of previouslyinfected subjects. Even more particularly, there has been no evidenceprovided heretofore of any methods of preventing infection through useof goose antibodies and subsequently reducing mortality in a populationat risk for infection by a transmittable viral disease. According to thepresent invention, there are provided methods of reducing mortality in apopulation of subjects by both treatment and prophylaxis through use ofgoose antibodies.

In addition to reducing mortality in a population of subjects, treatmentaccording to the present invention can be understood to relate tolessening or complete cessation of one or more symptoms associated withthe transmittable viral disease. Treatment can also include avoidingworsening of symptoms present at the time of first treatment andavoiding occurrence of further symptoms not present at the time of firsttreatment. Accordingly, in one embodiment, treatment can be effectedthrough preventing or limiting one or more symptoms associated withinfection by the transmittable viral disease.

Prevention of infection according to the present invention does notnecessarily mean completely preventing a subject from contracting atransmittable viral disease. Rather, a subject could contract thedisease after administration of the composition of the invention suchthat testing would indicate the presence of the virus in the subject,but the subject could be completely free of symptoms associated with thedisease. In such a case, even though a subject could test positive forthe disease, the absence of symptoms associated with the disease wouldindicate prevention. Accordingly, in one embodiment, the invention isdirected to a method of preventing active infection in a subject.Prevention could also be evidenced by a reduction in mortality in apopulation of subjects to which the composition of the invention hasbeen administered. In such a case, reduced mortality, when compared to apopulation not subject to administration of the inventive composition,would be indicative of prevention according to the invention.Preferably, prevention means complete absence of infection in a subjectto which the composition of the invention has been administered.

The present invention is particularly directed to methods andcompositions for treatment of transmittable viral diseases. As such, theinvention is capable of use in the treatment or prevention of a varietyof viral diseases. In some embodiments, the invention can be describedas relating to treatment or prevention of a viral disease that isrecognized as being treated, treatable, prevented, or preventable byadministration of antibodies against the transmittable viral disease.

In particular embodiments, the invention relates to treatment orprevention of infection by viruses in the family Flaviviridae. Infurther embodiments, the invention relates to treatment or prevention ofinfection by viruses in a specific genus encompassed by the Flaviviridaefamily. For example, the invention can relate to treatment or preventionof infection by viruses in the genus flavivirus. Flaviviruses share acommon size (i.e., about 40-65 nm), symmetry (i.e., enveloped,icosahedral nucleocapsid), and nucleic acid structure (i.e.,positive-sense, single stranded RNA having approximately 10,000-11,000bases). Flaviviruses have a (+) sense RNA genome and replicate in thecytoplasm of the host cells. The genome mimics the cellular mRNAmolecule in all aspects except for the absence of the poly-adenylated(poly-A) tail. This feature allows the virus to exploit cellularapparatus to synthesize both structural and non-structural proteinsduring replication. The cellular ribosome is crucial to the replicationof the flavivirus, as it translates the RNA, in a similar fashion tocellular mRNA, resulting in the synthesis of a single polyprotein. The(+) sense RNA genome of Flavivirus contain 5′ and 3′ untranslatedregions (UTRs). The 3′ UTRs are typically 0.3-0.5 kb in length andcontain a number of highly conserved secondary structures common to theflavivirus family. The present invention encompasses treatment relatedto tick-borne flaviviruses, mosquito-borne flaviviruses, andflaviviruses with no known arthropod vector. Specifically, the presentinvention encompasses treatment of flaviviruses, such as Gadgets Gullyvirus, Kadam virus, Kyasanur Forest disease virus, Langat virus, Omskhemorrhagic fever virus, Powassan virus, Royal Farm virus, Tick-borneencephalitis virus, Louping ill virus, Meaban virus, Saumarez Reefvirus, Tyuleniy virus, Aroa virus, Dengue virus, Kedougou virus,Cacipacore virus, Koutango virus, Japanese encephalitis virus, MurrayValley encephalitis virus, St. Louis encephalitis virus, Usutu virus,West Nile virus, Yaounde virus, Kokovera virus, Bagaza virus, Ilheusvirus, Israel turkey meningoencephalomyelitis virus, Ntaya virus,Tembusu virus, Zika virus, Banzi virus, Bouboui virus, Edge Hill virus,Jugra virus, Saboya virus, Sepik virus, Uganda S virus, Wesselsbronvirus, Yellow fever virus, Entebbe bat virus, Yokose virus, Apoi virus,Cowbone Ridge virus, Jutiapa virus, Modoc virus, Sal Vieja virus, SanPerlita virus, Bukalasa bat virus, Carey Island virus, Dakar bat virus,Montana myotis leukoencephalitis virus, Phnom Penh bat virus, and RioBravo virus.

In further embodiments, the invention can relate to treatment orprevention of infection by viruses in the family Orthomyxoviridae. Inspecific embodiments, the invention relates to treatment or preventionof infection by viruses in a specific genus encompassed by theOrthomyxoviridae. For example, the invention can relate to treatment orprevention by viruses in the genera influenzavirus A, influenzavirus B,or influenzavirus C. The term “influenza virus” as used herein canencompass viruses in any of the above-noted influenzavirus genera.

Influenzaviruses A, B and C are very similar in overall structure. Thevirus particle is 80-120 nanometers in diameter and usually roughlyspherical, although filamentous forms can occur (more commonly ininfluenza C), which can form cordlike structures up to 500 microns longon the surfaces of infected cells. Despite these varied shapes, theviral particles of all influenza viruses are similar in composition.These are made of a viral envelope containing two main types ofglycoproteins wrapped around a central core containing the viral RNAgenome and other viral proteins that package and protect this RNA. RNAtends to be single stranded but in special cases it is double stranded.The present invention encompasses treatment related to various types ofinfluenza viruses, including human influenza (or human flu), avianinfluenza (or bird flu), swine influenza, equine influenza, canineinfluenza, and cat flu. Hemagglutinin (HA) and neuraminidase (NA) arethe two large glycoproteins on the outside of influenza viral particles.HA is a lectin that mediates binding of the virus to target cells andentry of the viral genome into the target cell, while NA is involved inthe release of progeny virus from infected cells, by cleaving sugarsthat bind the mature viral particles. These proteins are targets forantiviral drugs, and they are antigens to which antibodies can beraised. Influenza A viruses are classified into subtypes based onantibody responses to HA and NA. These different types of HA and NA formthe basis of the H and N distinctions. The present invention encompassestreatment of influenza viruses having any recognized H and Nclassification. For example, the present invention encompasses treatmentof influenza viruses classified as H1N1, H1N2, H1N7, H2N2, H3N1, H3N2,H3N8, H4N8, H5N1, H5N2, H5N8, H5N9, H6N2, H6N5, H7N1, H7N2, H7N3, H7N4,H7N7, H8N4, H9N2, H10N7, H11N6, H12N5, H13N6, and H14N5.

In other embodiments, the invention can relate to treatment orprevention of infection by a variety of further viral diseases. Forexample, the invention can relate to treatment or prevention ofinfection from viruses of the family Herpesviridae. More specifically,the virus may be of one of the following genera: Iltovirus, Mardivirus,Simplexvirus, Varicellovirus, Cytomegalovirus, Muromegalovirus,Proboscivirus, Roselovirus, Lymphocryptovirus, Macavirus, Percavirus,and Rhadinovirus. In specific embodiments, the viral disease that can betreated or prevented according to the invention can comprise herpessimplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), varicellazoster virus (VZV), Eppstein-Barr virus, cytomegalovirus, roseolovirus,and Kaposi's sarcoma-associated herpesvirus (KSHV).

In further embodiments, the invention can relate to treatment orprevention of infection from viruses of the family Paramyxoviridae. Morespecifically, the virus may be of one of the following genera:Avulavirus (e.g., Newcastle disease virus), Henipavirus (e.g.,Hendravirus and Nipahvirus), Morbillivuris (e.g., Measles virus,Rinderpest virus, Canine distemper virus, Phocine distemper virus, Pestedes Petits ruminants virus), Respirovirus (e.g., Sendai virus, HumanParainfluenza viruses, and common cold viruses), Rubulavirus (e.g.,Mumps virus, Human Parainfluenza viruses, Simian Parainfluenza viruses,Menangle virus, Tioman virus, and Tuhokoviruses), Pneumovirus (e.g.,Human Respiratory Syncytial virus, and Bovine Respiratory Syncytialvirus), Metapneumovirus (e.g., Avian Pneumoirus and HumanMetapneumovirus), Fer-de-Lance virus, Nariva virus, TupaiaParamyxovirus, Salem virus, J virus, Mossman virus, and Beilong virus.

In other embodiments, the invention can relate to treatment orprevention of infection from viruses of the family Rhabdoviridae. Morespecifically, the virus may be of one of the following genera:Cytorhabdovirus (e.g., Lettuce necrotic yellows virus), Dichorhabdovirus(e.g., Orchid fleck virus), Ephemerovirus (e.g., Bovine ephemeral fevervirus), Lyssavirus (e.g., Rabies virus), Novirhabdovirus (e.g.,Infectious Hematopoietic necrosis virus), Nucleorhabdovirus (e.g.,Potato yellow dwarf virus), and Vesiculovirus (e.g., Vesicularstomatitis virus).

In certain embodiments, the invention can relate to treatment orprevention of encephalitis arising from viral infections. For example,the encephalitis treated or prevented according to the invention may beencephalitis arising from one or more of Herpes simplex, Varicellazoster virus, Rabies, HIV, and H5N1. In further examples, the inventioncan relate to treatment or prevention of an Arbovirus encephalitis(e.g., La Crosse encephalitis, California encephalitis virus, Japaneseencephalitis, St. Louis encephalitis, Equine encephalitis, Murray Valleyencephalitis, Tick-borne meningoencephalitis, Powassan encephalitis, andencephalitis arising from West Nile virus). Other viral encephalitidesthat can be treated or prevented according to the invention includeEastern equine encephalomyelitis virus, Venezuelan equineencephalomyelitis virus, and Western equine encephalomyelitis virus.

In other embodiments, the invention can relate to treatment orprevention of conditions recognized as viral hemorrhagic fevers (such asEbola, Marbug, Junin, Argentine, and Lassa). In certain embodiments, theinvention thus can relate to treatment or prevention of infection fromviruses of the family Arenaviridae, Filoviridae, Bunyaviridae, andFlaviviridae (which is already described above).

In relation to the family Arenaviridae, the virus may be of one of thefollowing genera: Ippy virus, Lujo virus, Lymphocytic choriomeningitisvirus, Mobala virus, Mopeia virus, Amapari virus, Chapare virus, Flexalvirus, Guanarito virus, Junin virus, Latino virus, Machupo virus,Oliveros virus, Parana virus, Pichinde virus, Pirital virus, Sabiavirus, Tacaribe virus, Tamiami virus, and Whitewater Arroyo virus. Inrelation to the family Filoviridae, the virus may be of the Marburgvirusgenus or the Ebolavirus genus. More specifically, the virus may includeZaire virus, Sudan ebolavirus, Reston ebolavirus, Cote d'Ivoireebolavirus (also known as Ivory Coast ebolavirus and Tai ebolavirus, andBundibugyo ebolavirus. In relation to the family Bunyaviridae, the virusmay be of one of the following genera: Hantavirus (e.g., Hantaan virus),Nairovirus (e.g., Dugbe virus), Orthobunyavirus (e.g., Bunyamweravirus), and Phlebovirus (e.g., Rift Valley fever virus).

In specific embodiments, non-limiting examples of further viral diseasesthat can be treated or prevented according to the invention includeHepatitis A, Hepatitis B, Ectocarpus Siliculosus Virus, and any otherviral diseases that are amenable to treatment and/or prevention viaadministration of antibodies. In some embodiments, the invention caneven relate to treatment or prevention of infection by diseases notpreviously recognized as being easily treated or prevented, such asHepatitis C and Human Immunodeficiency Virus.

In other embodiments, the invention can be characterized as providingfor treatment or prevention of infection by viruses recognized as RNAviruses. In specific embodiments, the methods and compositions of theinvention may be used in the treatment or prevention of viral diseasesclassified in any of the following families: Birnaviridae, Reoviridae(including Rotavirus), Coronaviridae [including Coronavirus and SevereAcute Respiratory Syndrome (SARS)], Picornaviridae (includingPoliovirus, the common cold virus, and Hepatitis A virus), Astroviridae,Caliciviridae (including Norwalk virus), Flaviviridae (as describedabove), Rhabdaviridae (including Rabies virus), Togaviridae (includingRubella virus, Ross River virus, Sindbis virus, and Chikungunya virus),Bornaviridae (including Borna disease virus), Filoviridae,Paramyxoviridae, Arenaviridae, Bunyaviridae, and Orthomyxoviridae. Suchclassification also includes viruses of the genus Hepevirus (includingHepatitis E virus), Deltavirus, and Nyavirus.

In other embodiments, the invention can be characterized as providingfor treatment or prevention of infection by viruses recognized as DNAviruses. In specific embodiments, the methods and compositions of theinvention may be used in the treatment or prevention of viral diseasesclassified in any of the following families: Myoviridae, Podoviridae,Adenoviridae, Asfarviridae, Herpesveridae, Hepadnaviridae,Papillomaviridae, Polyomaviridae, Poxviridae, Anelloviridae, andParvoviridae.

In still further embodiments, the invention can be characterized asproviding for treatment or prevention of infection by viral agents thatare cancer-causing viruses. Thus, the invention can be directed totreatment or prevention of viral oncogenes. Non-limiting examples ofsuch viruses include human papillomavirus, Hepatitis B, Hepatitis C,Epstein-Barr virus, Human T-lymphotropic virus, Kaposi'ssarcoma-associated herpesvirus, Merkel cell polyomavirus,

Compositions for use according to the invention can comprise goose serumcomprising antibodies to the specific viral disease. Such serum can beobtained or prepared by a variety of methods, and the serum can includea variety of components. As previously noted, the serum comprises gooseantibodies to the disease for which treatment or prevention is desired.The goose antibodies can be present naturally in the serum or can beincorporated into the serum as desired in the preparation of the serum.

In one preferred embodiment, the avian antibodies are naturally presentin the serum as obtained from a host. Accordingly, it is preferred forthe serum to comprise sera collected from one or more avian hosts.Preferably, the sera are collected from at least two avian hosts. Inspecific embodiments, as noted previously, the avian host can be agoose. Therefore, preparation of the inventive composition may befurther described herein in reference to obtaining sera from geese. Ofcourse, the invention also encompasses embodiments wherein the sera isobtained from further avians including, but not limited to ducks,turkeys, ostriches, chickens, and any further avians recognizable by oneof skill in the art as being useful in light of the further disclosureprovided herein.

Preferably, those avians of the biological family Anatidae, or commonlyknown as waterfowl, are preferred over other avian species. For example,the current highly pathogenic avian influenza (HPAI), H5N1, exhibitsvery high mortality, approaching or at 100%, to chickens and turkeys. Incontrast, waterfowl birds can be carriers of the H5N1 strain. In fact,various H5N1 strain variants can cause substantially reduced or nomortality in domestic waterfowl relative that observed in chickens. Theresistance of waterfowl is predicted to be due to the immunologicalsystem, and specifically antibodies, of the genus.

The avian hosts (e.g., geese) used for collecting the sera arepreferentially geese that have been infected by a naturally occurringstrain of the transmittable disease. Hosts having obtained a naturallyoccurring strain of the disease have been found to be particularly goodsources of antibodies effective in the inventive composition describedherein. The avian host can be a host that is actively infected or a hostthat has been previously infected but did not succumb to the disease.

The sera collected from the avian hosts can be used in the natural formor may be further processed or treated. For example, the sera arepreferably treated to substantially remove active forms of thetransmittable disease that may be present therein. In one embodiment,polyclonal antibodies are obtained from the serum. Such polyclonalantibodies may be used separate from the serum, used with a separateserum, or reintroduced into the same serum. The polyclonal antibodiescan be isolated using a variety of procedures, including chromatography,ammonium sulfate separation, molecular selection protocols, orcombinations thereof.

The serum used in a pharmaceutical composition and/or a method oftreatment according to the invention preferably exhibits a highneutralization titer for the avian antibodies. In preferred embodiments,the serum contains a high neutralization titer for goose antibodies. Asused herein, a neutralization titer is understood to mean a degree ofdilution at which a positive detection for a test component may still befound. Titer may be expressed in a variety of dilutions, and the use ofa specific dilution in describing the present invention should not beviewed as limiting the invention. By describing the serum as comprisinga high neutralization titer of avian antibodies, it is generally meantthat the neutralization titer of the serum for the avian antibodies ishigher that would be exhibited by the serum under normal conditions.According to one embodiment, normal conditions refers to conditionswherein the serum is obtained from a host that has not been infectedwith the disease for which antibodies are to be observed. According toanother embodiment, the serum neutralization titer can be consideredhigh titer if the neutralization titer is high than would be exhibitedif the serum was obtained from a host infected with a non-naturallyoccurring strain of the disease against which antibodies are to beobserved. In one specific embodiment, the neutralization titer of theserum is at least about 1:200. Preferably, the neutralization titer iseven higher. For example, in certain embodiment, it is preferable forthe neutralization titer of the serum for the avian antibodies to be atleast about 1:500, at least about 1:1,000, at least about 1:2,000, atleast about 1:3,000, or at least about 1:4,000. In certain embodiments,the neutralization titer for the serum is in the range of 1:320 to1:8,192, in the range of 1:512 to 1:8,192, in the range of 1:1,024 to1:8,192, or in the range of 1:2,048 to 1:4,096. Preferably,neutralization titer is evaluated in terms of polyclonal antibodies tothe disease to be observed.

Serum neutralization titer for protective antibodies can be a criticalfactor in the effectiveness of a treatment or prophylactic preparedusing the serum. Previous attempts to prepare formulations for treatingWest Nile Virus have centered on the use of the serum obtained fromhuman hosts. For example, Ben-Nathan et al. (J Infect Dis. 2003; 188:5-12) tested the efficacy of serum obtained from Israeli and U.S. humanhosts. They determined that immunoglobulin G (IgG) preparations fromIsraeli donors had an anti-WNV antibody titer of 1:1600 when evaluatedby ELISA. When evaluated by the plaque-reduction test, however, theantibody titer reported for the preparation was >1:80. Preparations madefrom U.S. blood donors were found to have a titer of only 1:10 whenmeasured by ELISA. One description of a plaque-reduction test isprovided by Yang et al., Journal of Immunological Methods 276, (2003):175-183, which is incorporated by reference in its entirety.

In light of these results, it was surprising, according to the presentinvention, to find that sera obtained from geese exhibited significantlyhigher neutralization titers for protective antibodies than exhibited bysera obtained from human hosts. Accordingly, compositions and methods ofthe present invention, which incorporate particularly highneutralization titer sera, would be expected to much more useful fortreating and preventing infection by viral diseases, such as West NileVirus. As one of skill in the art would recognize, neutralization titeris not a property that can be routinely optimized through manipulation.Rather, neutralization titer is a measurement of the neutralizingactivity of the antibodies present in the serum. Thus, the requisiteneutralization titer either is present or is not, and it is not possibleby routine optimization to convert a low neutralization titer serum intoa high neutralization titer serum.

Serum exhibiting a high neutralization titer, according to the presentinvention, against an indigenous transmittable virus can be obtainedusing a variety of methods that utilize avian hosts, particularly geese.In some embodiments, such a method can comprise exposing geese in anopen environment where the animals freely interact with a transmissionhost and with each other if a population is used. The preferred avian isone that can readily contract the transmittable virus of interest. Thetransmission host can be another avian capable of passing the disease onto host or can be a separate entity. For example, in the case of WestNile Virus, the transmission host is generally the mosquito. Testing hasindicated that polyclonal antibodies can be particularly obtained from apopulation of avian hosts where at least 3% of the avians previouslyinfected with the viral disease died, or in populations where theanimals already showed high neutralization titer to the transmittablevirus disease.

It has further been found that utilizing younger hosts for obtainingsera can be more effective, particularly for providing higherneutralization titer sera. For example, when geese are used as the hostfor obtaining the sera, obtaining sera from goslings rather thanbreeders has been found effective for obtaining sera with higherneutralization titers for protective antibodies.

Treatment according to the present invention can be carried out on avariety of subjects. In one embodiment, the invention comprises treatingor preventing infection by a transmittable viral disease in aviansubjects. While such treatment or prevention may be with any aviansubject, it is particularly useful with farm-raised avians, such asgeese, ducks, turkeys, ostriches, and chickens. Likewise, in otherembodiments, the invention is useful in treating or preventing infectionby a transmittable viral disease in mammalian subjects. Such mammaliansubjects can particularly include humans. In further embodiments, theinvention may be used in relation to further mammalian subjects, such asgoats, horses, rabbits, rats, mice, pigs, cat, dogs, and the like. Inanother preferred embodiment, any captive animal, such as exotic zooanimals, may be treated according to the invention prior to or afterbecoming infected with a transmittable viral disease. In particular, theinvention may be carried out with animals where a vaccine is noteconomically feasible or has not been shown to provide protection.

In light of the above, it becomes particularly clear that the inventionis particularly useful in relation to treating or preventing infectionin a subject by a transmittable viral disease. In certain embodiments,the present invention provides methods for treating or preventinginfection in a subject by administering to the subject goose antibodies(or suitable antibodies derived from another avian host) against thetransmittable viral disease. In particular embodiments, the methodcomprises administering to the subject an amount of a serum effectivefor treating or preventing infection by a transmittable viral disease,wherein the serum comprises goose antibodies against the transmittableviral disease.

Given the wide range of use associated with the above-noted methods, theadministration of the serum can take on a variety of schedules. Forexample, the administration of the serum could be carried out prior toinfection of the subject by the transmittable disease. Such a schedulewould be particularly effective in the prevention of infection by thedisease. In another embodiment, administration of the serum could becarried out at any point after infection of the subject by the disease.Such a schedule would be particularly effective as a treatment for thedisease. When treatment is indicated, such as by evidence of symptoms inthe subject common to the given disease, administration of the serum ispreferably given until symptoms are no longer evident. One quantifiablesymptom is the presence of virus neutralization titer levels in seracollected from the subject corresponding to the viral transmittabledisease. Preferably, the treatment according to the invention iseffective to reduce the virus neutralization titer levels in theinfected subject by 50% or more, and most preferably by at least 90%.

The serum may particularly be administered as part of a pharmaceuticalcomposition. As such, the compositions of the present invention cancomprise serum, together with one or more pharmaceutically acceptablecarriers therefore, and optionally, other therapeutic ingredients. By“pharmaceutically acceptable carrier” is intended a carrier that isconventionally used in the art to facilitate the storage,administration, and/or the healing effect of the agent. Carriers shouldbe acceptable in that they are compatible with any other ingredients ofthe composition and not harmful to the recipient thereof A carrier mayalso reduce any undesirable side effects of the agent. Such carriers areknown in the art. See, Wang et al. (1980) J. Parent. Drug Assn.34(6):452-462, herein incorporated by reference in its entirety.

Compositions of the present invention may include short-term,rapid-onset, rapid-offset, controlled release, sustained release,delayed release, and pulsatile release formulations, providing thecompositions achieve administration of the serum as described herein.See Remington's Pharmaceutical Sciences (18^(th) ed.; Mack PublishingCompany, Eaton, Pa., 1990), herein incorporated by reference in itsentirety.

Pharmaceutical compositions according to the present invention aresuitable for various modes of delivery, including oral, parenteral(including intravenous, intramuscular, subcutaneous, intradermal, andtransdermal), and inhalation. The most useful and/or beneficial mode ofadministration can vary, especially depending upon the condition of therecipient and the disease for which treatment or prevention is desired.In certain embodiment, administration can be by a combination of routes,for example, an initial oral dose followed by a schedule of injections.

The pharmaceutical compositions may be conveniently made available in aunit dosage form, whereby such compositions may be prepared by any ofthe methods generally known in the pharmaceutical arts. Generallyspeaking, such methods of preparation comprise combining (by variousmethods) an active agent, such as the serum comprising avian antibodies,with a suitable carrier or other adjuvant, which may consist of one ormore ingredients. The combination of the active ingredient with the oneor more adjuvants is then physically treated to present the compositionin a suitable form for delivery (e.g. shaping into a tablet or formingan aqueous suspension).

Compositions for parenteral administration include aqueous andnon-aqueous sterile injection solutions, which may further containadditional agents, such as anti-oxidants, buffers, bacteriostats, andsolutes, which render the compositions isotonic with the blood of theintended recipient. The compositions may include aqueous and non-aqueoussterile suspensions, which contain suspending agents and thickeningagents. Such compositions for parenteral administration may be presentedin unit-dose or multi-dose containers, such as, for example, sealedampoules and viles, and may be stores in a freeze-dried (lyophilized)condition requiring only the addition of the sterile liquid carrier, forexample, water (for injection), immediately prior to use.

It is contemplated that the serum comprising goose antibodies will beadministered to a subject (i.e., an avian or a mammal, preferably ahuman) in therapeutically effective amounts. That is, in an amountsufficient to effect treatment of a subject already infected by adisease or effect prevention of infection of the subject by the disease.In specific embodiments, an effective amount can be an amount effectiveto reduce mortality in a population infected by or at risk of infectionby the disease. The effective amount of the serum comprising gooseantibodies would be expected to vary according to the classification(e.g., avian or mammalian), weight, sex, age, and medical history of thesubject. Other factors which influence the effective amount may include,but are not limited to, the severity of the subject's condition, thespecific viral disease being treated, and the stability of the serum.Methods to determine efficacy and dosage are known to those skilled inthe art. See, for example, Isselbacher et al. (1996) Harrison'sPrinciples of Internal Medicine 13 ed., 1814-1882, herein incorporatedby reference.

In another embodiment of the invention, the pharmaceutical compositioncomprising the serum is administered intermittently. By “intermittentadministration” is intended administration of a therapeuticallyeffective dose of the serum, followed by a time period ofdiscontinuance, which is then followed by another administration of atherapeutically effective dose, and so forth. Administration of thetherapeutically effective dose may be achieved in a continuous manner,as for example with a sustained-release formulation, or it may beachieved according to a desired daily dosage regimen, as for examplewith one, two, three, or more administrations per day. By “time periodof discontinuance” is intended a discontinuing of the continuoussustained-released or daily administration of the serum comprising theavian antibodies. The time period of discontinuance may be longer orshorter than the period of continuous sustained-release or dailyadministration. During the time period of discontinuance, the antibodylevel in the subject may fall substantially below the maximum levelobtained during the treatment. The preferred length of thediscontinuance period depends on the concentration of the effective doseand the form of pharmaceutical composition used. The discontinuanceperiod can be at least 2 days, at least 4 days or at least 1 week. Inother embodiments, the period of discontinuance is at least 1 month, 2months, 3 months, 4 months or greater. When the pharmaceuticalcomposition is used as a vaccine, administration of the serum can beeffected through vaccination schedules that may be later determined mosteffective for achieving a maximum inoculation against a specifictransmittable viral disease.

The present invention, in addition to the above useful aspects, alsoprovides a diagnostic kit useful for detecting the presence of atransmittable viral disease in a sample. Generally, the sample will havebeen obtained from one or more subjects to determine the presence of anactive or dormant infection in the subject by a given disease. In oneembodiment, the diagnostic kit comprises goose polyclonal antibodiescapable of binding to antigens on the viral disease and a detectorcapable of detecting goose polyclonal antibodies bound to the antigens.The goose polyclonal antibodies used in the diagnostic kit can beobtained as otherwise described herein.

The detector used in the diagnostic kit of the invention can compriseany immunological detection means that use serum, polyclonal antibodies,or antibody fragments as the binding part of the detection. Examples ofsuch methods which would be useful according to the invention include,but are not limited to, ELISA, immunolocalization using taggedantibodies, Western blots, Ouchterlony double diffusion,immunoprecipitation, strip tests, or the like.

EXPERIMENTATION

The present invention is more fully illustrated by the followingexamples, which illustrate certain embodiments the present invention andare not to be construed as limiting.

Example 1 Preparation of High Neutralization Titer Serum for ViralProtective Bird Antibody

Antibody titer measured by serum neutralization (SN) assays wereperformed to provide analyses of the protective capabilities of thegoose antibody to viral infection over traditional ELISA assays thatmeasure binding affinity to viral epitopes. Such traditional methodshave shown discrepancies in the past. For example, in studies performedby Ben-Nathan et al. in obtaining antibodies from human subjects, theELISA titer was reported to be 1:1600; however, the functionalprotective capabilities were shown to be substantially lower and wereactually in the range of 1:80 to 1:320 (J Infect Dis 2003; 188: 5-12).

In the present study, geese of a variety of ages were exposed to WestNile Virus, sera were collected from the geese, and the sera of theinfected geese were tested using a sera microtiter neutralization plaqueassay to measure the usefulness of the sera for protecting cells fromviral infection and death. Briefly, a serial 2-fold dilution of goosesera (up to a dilution of 1:8192) were prepared in 96-well microtiterplates and 50 ul PFU of West Nile Virus were added. After incubation atroom temperature for 1 hr., 1×10⁴ Vero cells were added to the mixturesto test for plaque reduction and were incubated for seven days. Plaquereduction neutralizing titers were expressed as the reciprocal of thehighest dilution that gave 50% plaque reduction. Experiments indicatedthat goslings had a significantly higher titer for West Nile Virusantibodies than breeder stock geese, which are used for egg production.In fact, breeder stock geese exhibited a 4 fold lower titer for WestNile Virus antibodies than observed in the younger goslings.

Testing indicated that the sera obtained from the goslings exposed toWest Nile Virus had a SN titer of over 4000 by 3 weeks after exposure.The neutralization titer peaked to over 8000 between 35 days to 70 daysafter exposure and decreased to about 1000 by 90 days after exposure toWest Nile Virus. There was no evidence of reduced neutralization titerlevels in the ages examined. Surprisingly, in contrast to reports ofhyperimmune sera of adult human exposed to West Nile Virus, theneutralization titer of gosling sera was up to 100 fold higher than thatobserved in human sera (as evidenced by the Ben-Nathan et al. 2003 studyreferenced above). Accordingly, sera obtained from bird hosts infectedwith West Nile Virus proved to have much higher neutralization titer forprotective antibodies than sera obtained from human hosts.

Actual measured neutralization titer levels in sera collected fromgoslings infected with West Nile Virus, as described above, are providedbelow in Table 1. The age of the bird at the time of infection with WestNile Virus and the neutralization titer level at a specified number ofdays post-infection are provided.

TABLE 1 Titer Days After Titer Days After Days of Age Level InfectionDays of Age Level Infection 127 4,096 21 127 >8,192 69 128 4,096 22 1212,048 69 128 4,096 22 127 4,096 69 120 4,096 25 125 2,048 73 132 4,09626 132 4,096 73 120 4,096 26 120 4,096 75 123 4,096 26 125 2,048 75 1254,096 28 120 4,096 76 126 4,096 29 126 4,096 76 135 >8,192 30 126 2,04876 127 4,096 30 126 2,048 76 124 4,096 30 103 4,096 76 132 4,096 32 1334,096 77 133 4,096 33 135 4,096 77 134 4,096 33 127 4,096 77 128 >8,19234 128 4,096 77 132 >8,192 39 125 4,096 78 128 4,096 40 140 4,096 81127 >8,192 46 125 4,096 82 125 4,096 51 144 4,096 84 123 4,096 52 1254,096 84 133 >8,192 52 113 2,048 84 134 >8,192 53 126 4,096 85127 >8,192 53 114 2,048 85 133 >8,192 54 126 4,096 86 141 >8,192 60 1171,024 88 127 4,096 64 117 2,048 88 127 4,096 65 120 2,048 88 133 4,09666 126 1,024 90 130 4,096 66 114 1,024 94 134 4,096 67 114 1,024 94126 >8,192 68 153 1,024 95 126 4,096 68

Example 2 Evaluation of Goose Antiserum for Presence of West Nile Virus

Goose antiserum was examined for the presence of West Nile Virus RNA byRT-PCR analysis. DNA was amplified from the prepared RNA in aPerkin-Elmer Model 480 thermal cycler. Primers were designed to map theconserved sequences of the polyprotein gene (West Nile Virus OligoDetect Kit, WNV Primer Mix [Part No. 5653], Chemicon International,California). The RT-PCR was performed with the QIAGEN one-step RT-PCRkit (QIAGEN, Valencia, Calif.) by using 5 μl of RNA and 0.3 μM of eachprimer in a 50 μl total reaction volume following the manufacture'sprotocol. When the PCR mixture was complete, the samples were overlaidwith two drops of molecular biology grade mineral oil. All previousmanipulations were performed in a Nuaire biological safety cabinet ModelNU 425-400. The following cycling times and temperatures were used: cDNAsynthesis; 50° C. for 30 minutes, 94° C. for 15 minutes followed by 40cycles of 94° C. for 1 minute, 57° C. for 30 seconds and 72° C. for 1minute followed by 72° C. for 15 minutes and 4° C. storage. FollowingPCR amplification of the DNA samples, the products were separated on 3%submerged agarose gels by electrophoresis. The separated products werevisualized by staining with ethidium bromide and electronicallyphotographed using UVP GDS8000 Gel Documentation System (Ultra VioletProducts).

Goose antiserum was examined for the presence of West Nile Virus RNA byRT-PCR analysis performed using Chemicon West Nile Virus OLIGODETECT® onantiserum samples. RNA was isolated from goose antisera samples usingthe QIAamp Viral RNA kit (available from Qiagen) following its suggestedprotocol. The protocol utilized AVL/carrier RNA addition to sera samplefollowed by application to a QIAamp spin column. After washing unboundmaterial with AW2, the RNA was eluted using AVE buffer and collected bycentrifugation. For RT-PCR analysis West Nile Primer Mix was added insuggested amounts to Qiagen One-Step RT-PCR Enzyme and sample, whichincluded either RNA isolated from antisera or positive control West NileVirus RNA included in kit. RT-PCR was performed as recommended by kitand subjected to agarose gel analysis.

The positive control West Nile Virus RNA sample exhibited the presenceof the appropriate approximately 100 base pair PCR product, however noPCR products were observed in either the negative control or antiserasamples. This observation ruled out potential artifacts caused by thepresence of West Nile Virus particles in goose antiserum acting as avaccine rather therapeutic agent.

Example 3 Purification of Goose Antibodies to West Nile Virus

Twenty liters of sera collected from geese infected with West Nile Viruswas irradiated for 67 minutes/300 ml aliquots to eliminate any residualvirus present in the sera, and the samples were examined by polymerasechain reaction (PCR) to ensure that the sera was virus free. Theantibody fraction of the sera was purified by density centrifugation,dialyzed to remove gradient, and concentrated to approximately 3 timesthe original protein concentration. Purity of the goose antibody wasestablished using RT-PCR analysis. All preparations were greater than1:4000 determined by a microtiter plaque neutralization assay.

Example 4 Detection of West Nile Virus in Sick Birds using VECTEST®

VECTEST® (Medical Analysis Systems) is recognized by the Center forDisease Control (CDC) as an effective rapid screening test for thedetection of West Nile Virus. 39 geese exhibiting advanced West NileVirus symptoms (e.g., lethargy, staggering, or blindness) were testedfor the presence of the virus using VECTEST®. In this “on farm” use,only 5 geese of the 39 geese tested showed a positive VECTEST® response,ranging from +1 to +3 on the test scale. In contrast, the majority oftested geese were found to be positive for West Nile Virus when testedusing RT-PCR. Within 7 days, all 5 of the birds testing positive by theVECTEST® died; however, 19 of the 34 birds testing negative by theVECTEST® died within the same period. Accordingly, VECTEST® analyses ofserum failed to detect a positive response in most of the birds testedaccording to the kit instructions.

The brains of selected geese were examined for the presence of West NileVirus and confirmed to be positive by histopathology for WNV lesions andby molecular PCR diagnostics by the Veterinary Diagnostic Laboratory atthe University of Minnesota. The VECTEST® is a monoclonal antibody testbased on a Saint Louis Encephalitis antigen panel and appears not to besufficiently reactive to indigenous West Nile Virus. In contrast, thegoose polyclonal antibodies reacted strongly to the indigenous West NileVirus antigens present in the indigenous population and unexpectedlyoffer higher reactivity than the monoclonal antibodies used in theVECTEST®.

Example 5 Mortality Rate of Avians Treated with Antiserum Prior to Onsetof West Nile Virus Disease

Prior to any evidence of increased mortality due to natural infection byWest Nile Virus, 6-10 week old goslings were treated by a singlesubcutaneous injection of 3 milliliters goose antisera according to theinvention per gosling. The antisera was negative for West Nile Virus RNAand had a neutralization titer of ≧1:2,000. The antisera treated groupincluded 4705 males and 5095 females. As a control, an untreated groupincluding 5462 males and 7536 females was also evaluated. All animalswere continuously exposed to natural infection by the West Nile Virusthroughout the course of a six week period.

Deaths among the birds injected with the antisera and the control groupwas recorded over a 19 day period beginning one day after immunizationwhen the mortality rates became elevated in control group. The percentmortality rates were calculated after first subtracting the averagebackground mortality rate observed in periods when West Nile Virusoutbreak was not observed. The percent mortality rate for treated anduntreated female and male birds is shown below in Table 2. The overallmortality rate decreased by approximately 60% to 80% in those goslingstreated with antiserum. This results show that injection of antiserumcan provide an effective control for reducing mortality rates due toWest Nile Virus when given prior to onset of a natural outbreak of thedisease in a population. RT-PCR analysis of the antiserum for West NileVirus RNA indicated antibodies residing in the antiserum were effectiveagents in reducing mortality.

TABLE 2 % Mortality % Mortality % Reduced Gender in Untreated in TreatedMortality Rate Male 5.46 1.34 75% Female 2.56 1.10 57%

Example 6 Mortality Rate After Onset of West Nile Virus Disease ofAvians Treated with Antiserum

In another study, goslings showing signs of West Nile Virus infection asjudged by a higher mortality were given a single injectionsubcutaneously with 3 milliliters of goose antisera. The antisera wasnegative was West Nile Virus RNA and had a neutralization titer level≧1:2000. The antisera treated group included 2463 males and 2379 femalesand an untreated control group included 5256 males and 7419 females.Deaths among the injected goslings and the control group were recordedover a 13 day period after the start of injection (such periodcorresponding to a period when mortality rates were elevated). Thepercent mortality rates were calculated after first subtracting theaverage background mortality rate observed in periods when West NileVirus outbreak was not observed. The percent mortality rate for treatedand untreated female and male birds is provided below in Table 3. Theoverall mortality rate in goslings treated with antiserum was decreasedby 57% to 68%. These results show that injection of antiserum, andspecific antibodies therein, provided an effective treatment forreducing mortality rates due to West Nile Virus after the naturaloutbreak of the disease in a population.

TABLE 3 % Mortality % Mortality % Reduced Gender in Untreated in TreatedMortality Rate Male 3.52 1.50 57% Female 3.94 1.26 68%

Example 7 Evaluation of Prophylaxis Against West Nile Virus in a LargePopulation of Geese Via Administration of Therapeutic Antibodies to aSubset of the Population

It is unclear whether in a goose flock affected by West Nile Virus thedisease is spread by transmission via animal to animal in addition tonaturally transmission by the original host (i.e., mosquitoes). A studywas preformed to determine if the treatment of a sufficient segment ofanimal population is effective to prevent the spread of the diseasewithin a large population of the entire flock, by potentially eitherreducing the transmission via animal to animal or reducing the viralpool for mosquitoes. Two sites (approximately 10 miles apart) wereselected for testing, both sites being known to have previously had asimilar mortality rate in geese due to natural infection by West NileVirus. At site 1, no geese were treated. At site 2, approximately 65% ofthe geese were treated with goose sera or antibodies.

WNV was observed at site 1 approximately 10 days prior to firstobservance at site 2. At site 1, the mortality rate associated with WestNile Virus infection was approximately 13.5% of the population over 25days. In contrast, at site 2, in antibody treated geese, the mortalityrate was 1.56% over the same day period corresponding to an 8 folddecrease in mortality relative to site 1. Surprisingly, in 35% of thegeese at site 2 that were not injected with sera, the mortality was alsosubstantially decreased relative to site 1; nevertheless, the mortalityrate in untreated geese at site 2 was still higher than those treatedwith the antibody. This suggests a benefit to untreated animals in alarger population arising from treatment of a subset of the population.Results are shown in Table 4.

TABLE 4 Fold Reduction in Treatment Farm Site Mortality % MortalityUntreated 1 13.58% — Antiserum Treated 2 1.56% 8.70 Untreated 2 3.02%4.39

Example 8 Toxicity and Longevity Studies in Mammals of Goose Antibodiesto West Nile Virus

Ten young adult mice were injected intramuscularly with 0.2 ml purifiedgoose antibodies to West Nile Virus, 10 separate young adult mice wereinjected intramuscularly with 0.4 ml purified goose anti-WNV antibodies,and 10 separate age-matched control mice were injected intramuscularlywith saline. All mice were observed for the first 24 hours and daily for3 weeks for adverse clinical symptoms including changes in food andwater consumption, wasting, and grooming. Neither acute nor chronicsymptoms were detected in any of the antibody treated mice.

At 3 weeks post-injection, all 30 of the mice were euthanized andexamined for gross anatomical changes with none detected. All spleensand livers were removed and analyzed histologically. No inflammation wasnoted in any of the experimental mice, and no difference was detectedbetween the antibody treated and control mice. There was no indicationthat there was any adverse reaction with the introduction of gooseantibodies into the mice.

Example 9 Efficacy of Goose Antibodies Against West Nile Virus inHamsters

To determine if goose antibodies to West Nile Virus would be effectivein mammals, the golden hamster model of WNV infection was utilized. Thegolden hamster model is discussed by Tesh et al., Persistent West NileVirus Infection in the Golden Hamster: Studins on its Mechanism andPossible Implications for Other Flavivirus Infections, The Journal ofInfectious Diseases (2005), 192:287-295, and Xiao et al., West NileVirus Infection in the Golden Hamster (Mesocricetus auratus): a Modelfor West Nile Encephalitis, Emerging Infectious Diseases (2001),7(4):714-721, both of which are incorporated herein by reference intheir entirety.

In the present test, ten hamsters were injected with purified gooseantibodies to West Nile Virus, and 10 control hamsters received saline(the day of injection being day 0). On day 1 all 20 hamsters wereinfected with 10^(3.2) PFU (plaque forming units) of WNV-Iowa strain.

The antibody-treated group and the saline group each divided into twogroups of five hamsters, the groups being orbitally bled either on days1 and 3 or days 2 and 4. WNV neutralization titer was determined in 1:5dilutions of sera. WNV was detectable in the control group by day 1 andincreased until the third day when the neutralization titer leveled off(see Table 5 below). Eight of the 10 hamsters from the saline groupshowed a positive WNV neutralization titer at the level tested. Incontrast to the saline treated animals, none of the hamsters receivingthe goose antibodies showed any functional virus at the lowest dilution10⁻¹.

TABLE 5 Average WNV-Ia Titer/mL Study Group Day 1 Day 2 Day 3 Day 4Saline Group Group 1 2.01 × 10² 1.67 × 10⁵ Group 2 3.34 × 10⁴ 9.19 × 10⁴Antibody Treated Group 1 ND ND Group 2 ND ND ND—Not Detected

To detect long term effects of the viral infection beyond the four-daytest described above, the hamsters were monitored for the next 11 daysfor clinical signs of West Nile Virus, including lethargy, wasting, anddeath. The hamsters receiving only saline showed 60% overall mortalityover days 4 through 11. Mortality was evidenced by natural death oreuthanization in light of viral effects. Complete mortality rates forboth groups are shown below in Table 6.

TABLE 6 Study Group Died or Euthanized Saline Group Group 1 2/5 Group 24/5 Antibody Treated Group 1 0/5 Group 2 0/5

As evidenced by the results provided in Table 5 and Table 6, gooseantibodies were shown effective for preventing infection by West NileVirus in mammals

Example 10 Preparation of High Neutralization Titer Serum Against AvianInfluenza

To test production of therapeutic antibody to avian influenza, an avianinfluenza virus strain was produced in eggs as a vaccine stock. A stocksample of H3N2 was obtained from ATCC (VR-777) culture collection andused as a viral stock for injection into waterfowl eggs. Two lines, P2SMand JMOP, of goose embryos were used for virus production. Goose embryosat 11 to 17 days of incubation were candled for viability prior to viralinjection. Holes were drilled at positions on egg that provided accessto either the air sac or chorioallantoic membranes. Approximately 10 to100 ul of virus stock solution was placed in the air sac or injectedinto the chorioallantoic membrane. The hole was sealed using glue andreturned in the upright position into an incubator. The eggs weremonitored for viability by candling.

After 3 to 6 days, approximately 0.5-1.0 ml of allantoic fluid werecollected from the allantoic cavity of the goose embryos. RNA wasextracted from the samples and analyzed according to the protocolrecommended in the RT-PCR kit (available from Qiagen) used for detectionof H3N2 virus. Briefly, 500 ul of allantoic fluid were mixed with 500 ulof RLT buffer. From this, 700 ul was applied to a RNeasy column andmicrofuged for 15 sec and repeated with the remaining sample. 700 ul ofBuffer RW1 was applied and the column was microfuged for 15 sec. Next,500 ul of RPE was similarly applied and microfuged and repeated. Toelute bound RNA, 30 to 50 ul of RNase free water was added andmicrofuged for 15 sec and the sample collected for RT-PCR.

RT-PCR was performed using H3N2 primers obtained from Integrated DNATechnologies, Inc. (Coralville, Iowa). The primer set included a forwardprimer, M2F, and a reverse primer, M253R. RT PCR was performed accordingto the Influenza A virus protocol by Fouchier et al. (J. Clin.Microbiology 38, 2000), which is incorporated herein by reference.Briefly, RT-PCR conditions were maintained for 30 min at 42° C. and 4min at 95° C. followed by 40 cycles of 1 min at 95° C., 1 min at 45° C.and 3 min at 72° C. Approximately 15 ul of nucleotide sample was addedto a reaction containing 5 ul of each primer and mixed with RT-PCRbuffer containing TAQ enzyme and dNTP. Samples of RT-PCR were analyzedby agarose electrophoresis and ethidium bromide staining.

In control eggs (mock injected, or eggs injected with virus butharvested after 3 hours), no virus was detected by the RT-PCR. Incontrast, H3N2 virus was found to be produced in 8 of 10 goose embryos.Embryos of both goose strains were shown to produce virus. Highest virusproduction was exhibited upon injection into the allantoic sac comparedto the air sac.

In the event that multiple strains of the transmittable virus arepresent, two or more strains can be inoculated individually intoresistant avian embryos and the allantoic are pooled to provide broaderprotection to strain variants. For common influenza vaccine typicallythree predominant strains from the past and/or during the present yearare used. After the allantoic fluids are pooled, a number of methodshave been used to simplify the recovery of the virus or viral productsfrom the allantoic fluids. Examples of such methods can be found in thefollowing, all of which are incorporated by reference herein: U.S. Pat.No. 3,627,873; U.S. Pat. No. 4,000,527; U.S. Pat. No. 3,316,153; U.S.Pat. No. 4,724,210; and U.S. Pat. No. 3,962,421.

The isolated H3N2 influenza viral particles are attenuated by a numberof methods to those skilled in the art that inactivate the viral nucleicacid or disrupt key viral coat elements critical in viral infection or acombination of methods. The attenuated viral particles are injected oneor more times into goslings for a period sufficient to produce an immuneresponse, typically 3 to 10 weeks. The sera are then harvested from theanimals and the sera are tested using a sera microtiter neutralizationplaque assay to measure their usefulness in protecting Vero cells fromviral infection. Upon demonstrating high neutralization titer goosesera, antibodies are isolated from sera by density centrifugation. Theantibodies are dialyzed to remove gradient and are concentrated toapproximately 3 times the original protein concentration.

The goose purified antibodies to avian influenza are tested in animalsand are found to be effective for preventing infection or preventing orlimiting one or more symptoms associated with infection by avianinfluenza.

Example 11 Generation of High Neutralization Titer Goose Sera AgainstH1N1 Influenza Virus

The ability of geese to generate a strong antibody response was testedby injection with inactivated H1N1 influenza virus (A/Cal/04/2009).Geese, eight with equal gender diversity, were injected with 500microliters of a solution of inactivated H1N1 virus and CompleteFruend's adjuvant (1:1 ratio). Each goose received 250 micrograms ofinactivated virus. The geese received booster immunizations of thecorresponding inactivated virus concentration in Incomplete Fruend'sadjuvant at 2, 4, 6, and 8 weeks after the initial injection. The geesewere bled prior to the initial immunization and with each subsequentimmunization. Immunization elicited a very robust antibody response inall of the geese by week 6 (corresponding to the 3^(rd) boosterimmunization). The response in all geese exceeded 30,000hemagglutination inhibition assay titers (HA-I). The peak HA-I titers atweek 6 (observed in half of the geese tested) were >120,000. Testingthus demonstrated that geese can be induced to generate antibodies toH1N1, a finding that would be expected to extend across the family ofinfluenza viruses. Testing also showed that the anti-influenza antibodyresponse in geese is generated very quickly and reaches exceptionallyhigh titers within a relatively short time after the initialintroduction of the virus.

Example 12 Epitope Mapping of Goose-Anti-WNV Antibodies as Evidence ofCross-Family Treatment Ability

Viruses in the Flaviviridae family (e.g. West Nile Virus (WNV), denguevirus, and Yellow Fever Virus) are relatively conserved, especially withrespect to their non-structural proteins. Testing was carried out toevaluate the possibility that goose antibodies to one member of theFlaviviridae family could provide cross-treatment capability with othermembers of the same family and to identify characteristics common to theFlaviviridae family indicating that goose antibodies would be expectedto provide effective treatments in a significant proportion of theviruses in the family. Epitope-mapping of goose anti-WNV antibodies wascarried out using JPT Replitope™ materials (available from JPT PeptideTechnologies GmbH, Berlin, Germany). Test results indicated that twoepitopes recognized by the IgY from WNV infected geese matched twodengue virus epitopes mapped by other groups. The overlapping WNVepitope and dengue virus epitope were located on their respectivenon-structural 1 (NS1) proteins (Falconar, A. K., P. R. Young, et al.(1994). “Precise location of sequential dengue virus subcomplex andcomplex B cell epitopes on the nonstructural-1 glycoprotein.” Arch Virol137(3-4): 315-326). Monoclonal antibodies specific for dengue virus NS1have been shown to be protective against later dengue challenge. Thesecond overlapping epitope between WNV and dengue virus was found ontheir NS3 proteins. NS3 has a number of enzymatic activities and isconsidered to be primarily intracellular in both viruses. Despite this,we identified the highest number (i.e., 20) IgY WNV-specific-epitopes onNS3 compared to all proteins assayed, and this showed the strongestbinding of all of the IgY epitopes identified on WNV. For dengue virus,NS3 has been identified as mainly having T-cell epitopes—particularlyfour core NS3 T-cell epitopes (Brinton, M. A., I. Kurane, et al. (1998).“Immune mediated and inherited defenses against flaviviruses.” ClinDiagn Virol 10(2-3): 129-139). When this data was compared to theresults from the WNV sera, not only was NS3 relatively conserved acrossdengue virus and WNV, but also the epitope that produced the highestmedian fluorescence intensity (MFI) when bound to IgY overlapped withone of the four strong T-cell epitopes in WNV identified by Brinton etal. This strongly suggests that goose antibodies to WNV may be effectiveacross the entire Flaviviridae family. These results further suggestthat the ability of geese to produce antibodies to WNV and the abilityof goose anti-WNV antibodies to be an effective treatment in mammalswould extend to other members of the Flaviviridae family such that geesewould be expected to produce antibodies to other members of the family,and that the produced antibodies would be expected to provide effectivetreatments in mammals.

Many modifications and other embodiments of the inventions set forthherein will come to mind to one skilled in the art to which theseinventions pertain having the benefit of the teachings presented in theforegoing descriptions. Therefore, it is to be understood that theinventions are not to be limited to the specific embodiments disclosedand that modifications and other embodiments are intended to be includedwithin the scope of the appended claims. Although specific terms areemployed herein, they are used in a generic and descriptive sense onlyand not for purposes of limitation.

1. A method of treating a mammal infected with a virus in theFlaviviridae family comprising administering to the mammal an amount ofa serum comprising polyclonal goose antibodies against a virus in theFlaviviridae family.
 2. The method of claim 1, wherein the mammal isselected from the group consisting of goats, horses, rabbits, rats,mice, pigs, and humans.
 3. The method of claim 1, wherein the virusinfecting the mammal is a flavivirus.
 4. The method of claim 1, whereinthe flavivirus is selected from the group consisting of Gadgets Gullyvirus, Kadam virus, Kyasanur Forest disease virus, Langat virus, Omskhemorrhagic fever virus, Powassan virus, Royal Farm virus, Tick-borneencephalitis virus, Louping ill virus, Meaban virus, Saumarez Reefvirus, Tyuleniy virus, Aroa virus, Dengue virus, Kedougou virus,Cacipacore virus, Koutango virus, Japanese encephalitis virus, MurrayValley encephalitis virus, St. Louis encephalitis virus, Usutu virus,West Nile virus, Yaounde virus, Kokovera virus, Bagaza virus, Ilheusvirus, Israel turkey meningoencephalomyelitis virus, Ntaya virus,Tembusu virus, Zika virus, Banzi virus, Bouboui virus, Edge Hill virus,Jugra virus, Saboya virus, Sepik virus, Uganda S virus, Wesselsbronvirus, Yellow fever virus, Entebbe bat virus, Yokose virus, Apoi virus,Cowbone Ridge virus, Jutiapa virus, Modoc virus, Sal Vieja virus, SanPerlita virus, Bukalasa bat virus, Carey Island virus, Dakar bat virus,Montana myotis leukoencephalitis virus, Phnom Penh bat virus, and RioBravo virus.
 5. The method of claim 1, wherein the serum comprisespolyclonal goose antibodies against a virus in the flavivirus genus. 6.The method of claim 5, wherein the virus is selected from the groupconsisting of Gadgets Gully virus, Kadam virus, Kyasanur Forest diseasevirus, Langat virus, Omsk hemorrhagic fever virus, Powassan virus, RoyalFarm virus, Tick-borne encephalitis virus, Louping ill virus, Meabanvirus, Saumarez Reef virus, Tyuleniy virus, Aroa virus, Dengue virus,Kedougou virus, Cacipacore virus, Koutango virus, Japanese encephalitisvirus, Murray Valley encephalitis virus, St. Louis encephalitis virus,Usutu virus, West Nile virus, Yaounde virus, Kokovera virus, Bagazavirus, Ilheus virus, Israel turkey meningoencephalomyelitis virus, Ntayavirus, Tembusu virus, Zika virus, Banzi virus, Bouboui virus, Edge Hillvirus, Jugra virus, Saboya virus, Sepik virus, Uganda S virus,Wesselsbron virus, Yellow fever virus, Entebbe bat virus, Yokose virus,Apoi virus, Cowbone Ridge virus, Jutiapa virus, Modoc virus, Sal Viejavirus, San Perlita virus, Bukalasa bat virus, Carey Island virus, Dakarbat virus, Montana myotis leukoencephalitis virus, Phnom Penh bat virus,and Rio Bravo virus.
 7. The method of claim 1, wherein the virus thatthe polyclonal goose antibodies are against is the same virus infectingthe mammal.
 8. The method of claim 1, wherein the virus that thepolyclonal goose antibodies are against is different from the virusinfecting the mammal.
 9. The method of claim 1, wherein said treatingcomprises preventing or limiting one or more symptoms associated withinfection by the virus.
 10. The method of claim 1, wherein the serumexhibits a neutralization titer of at least about 1:200 when evaluatedaccording to a plaque reduction test.
 11. The method of claim 10,wherein the serum exhibits a neutralization titer of at least about1:500 when evaluated according to a plaque reduction test.
 12. Themethod of claim 10, wherein the serum exhibits a neutralization titer ofat least about 1:1000 when evaluated according to a plaque reductiontest.
 13. The method of claim 1, wherein the mammal has not beende-sensitized to goose antibodies prior to said step of administeringthe serum comprising the polyclonal goose antibodies against the virusin the Flaviviridae family.
 14. The method of claim 1, wherein saidadministering comprises a route of administration selected from thegroup consisting of injection, inhalation, oral administration, andcombinations thereof.
 15. A method of treating a mammal infected with aninfluenza virus comprising administering to the mammal an amount of aserum comprising polyclonal goose antibodies against an influenza virus.16. The method of claim 15, wherein the mammal is selected from thegroup consisting of goats, horses, rabbits, rats, mice, pigs, andhumans.
 17. The method of claim 15, wherein the virus infecting themammal is a virus in the genus influenzavirus A.
 18. The method of claim17, wherein the influenza virus is selected from the group consisting ofH1N1, H1N2, H1N7, H2N2, H3N1, H3N2, H3N8, H4N8, H5N1, H5N2, H5N8, H5N9,H6N2, H6N5, H7N1, H7N2, H7N3, H7N4, H7N7, H8N4, H9N2, H10N7, H11N6,H12N5, H13N6, and H14N5.
 19. The method of claim 15, wherein the serumcomprises polyclonal goose antibodies against a virus in the genusinfluenzavirus A.
 20. The method of claim 19, wherein the virus isselected from the group consisting of H1N1, H1N2, H1N7, H2N2, H3N1,H3N2, H3N8, H4N8, H5N1, H5N2, H5N8, H5N9, H6N2, H6N5, H7N1, H7N2, H7N3,H7N4, H7N7, H8N4, H9N2, H10N7, H11N6, H12N5, H13N6, and H14N5.
 21. Themethod of claim 15, wherein the virus that the polyclonal gooseantibodies are against is the same virus infecting the mammal.
 22. Themethod of claim 15, wherein the virus that the polyclonal gooseantibodies are against is different from the virus infecting the mammal.23. The method of claim 15, wherein said treating comprises preventingor limiting one or more symptoms associated with infection by the virus.24. The method of claim 15, wherein the serum exhibits a neutralizationtiter of at least about 1:200 when evaluated according to a plaquereduction test.
 25. The method of claim 24, wherein the serum exhibits aneutralization titer of at least about 1:500 when evaluated according toa plaque reduction test.
 26. The method of claim 24, wherein the serumexhibits a neutralization titer of at least about 1:1000 when evaluatedaccording to a plaque reduction test.
 27. The method of claim 15,wherein the mammal has not been de-sensitized to goose antibodies priorto said step of administering the serum comprising the polyclonal gooseantibodies against the influenza virus.
 28. The method of claim 15,wherein said administering comprises a route of administration selectedfrom the group consisting of injection, inhalation, oral administration,and combinations thereof.
 29. A pharmaceutical composition comprising aserum effective for treatment or prophylaxis of a viral infection in amammal arising from a virus in the Flaviviridae family, the serumcomprising polyclonal goose antibodies against a virus in theFlaviviridae family, the serum exhibiting a neutralization titer for theantibodies of at least about 1:500 when evaluated according to a plaquereduction test.
 30. A pharmaceutical composition comprising a serumeffective for treatment or prophylaxis of a viral infection in a mammalarising from an influenza virus, the serum comprising polyclonal gooseantibodies against an influenza virus, the serum exhibiting aneutralization titer for the antibodies of at least about 1:500 whenevaluated according to a plaque reduction test.